
浏览全部资源
扫码关注微信
1.南京中医药大学 第一临床医学院,南京 210046
2.南京中医药大学 附属医院,江苏省中医院,南京 210029
史露露,硕士,从事不孕症的临床及基础研究,E-mail:648818013@qq.com
* 周惠芳,博士,教授,博士生导师,从事不孕症的临床及基础研究,E-mail:zhouhuifang2011301@163.com
收稿日期:2021-02-18,
网络出版日期:2021-06-23,
纸质出版日期:2021-08-20
移动端阅览
史露露,周惠芳.补肾助孕方对黄体功能不全模型大鼠卵巢内质网应激的调节[J].中国实验方剂学杂志,2021,27(16):84-89.
SHI Lu-lu,ZHOU Hui-fang.Regulation of Bushen Zhuyun Prescription on Ovarian Endoplasmic Reticulum Stress in Rats with Luteal Phase Defect[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(16):84-89.
史露露,周惠芳.补肾助孕方对黄体功能不全模型大鼠卵巢内质网应激的调节[J].中国实验方剂学杂志,2021,27(16):84-89. DOI: 10.13422/j.cnki.syfjx.20211692.
SHI Lu-lu,ZHOU Hui-fang.Regulation of Bushen Zhuyun Prescription on Ovarian Endoplasmic Reticulum Stress in Rats with Luteal Phase Defect[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(16):84-89. DOI: 10.13422/j.cnki.syfjx.20211692.
目的
2
探讨补肾助孕方对米非司酮诱导的黄体功能不全(LPD)模型大鼠卵巢内质网应激(ERS)的调控机制。
方法
2
使用随机数表法将50只SD大鼠分为空白组,模型组,地屈孕酮(0.02 g·kg
-1
)组,补肾助孕方低(0.08 g·kg
-1
)、高剂量(0.24 g·kg
-1
)组。使用蛋白免疫印迹法(Western blot)及实时荧光定量聚合酶链式反应(Real-time PCR)检测各组大鼠卵巢免疫球蛋白重链结合蛋白(BIP),蛋白激酶R样内质网激酶(PERK),肌醇必需酶-1(IRE-1),活性转录因子6(ATF6),C/EBP同源蛋白(CHOP)蛋白和mRNA表达水平,酶联免疫吸附测定法(ELISA)检测各组大鼠血清孕酮(P)和雌二醇(E
2
)水平。
结果
2
与空白组比较,模型组BIP,PERK,CHOP蛋白表达显著升高(
P
<
0.01),PERK及CHOP mRNA表达明显下降(
P
<
0.05);而IRE-1,ATF6蛋白表达,IRE-1,BIP,ATF6 mRNA表达,血清E
2
,P水平差异无统计学意义。与模型组比较,地屈孕酮组CHOP蛋白表达显著降低(
P
<
0.01),而PERK,IRE-1,BIP,ATF6蛋白表达,PERK,IRE-1,BIP,ATF6,CHOP mRNA表达及血清E
2
,P水平差异无统计学意义;补肾助孕方低剂量组CHOP蛋白表达显著降低(
P
<
0.01),CHOP mRNA表达水平明显升高(
P
<
0.05),而PERK,IRE-1,BIP,ATF6蛋白和mRNA表达及血清E
2
,P水平差异无统计学意义;补肾助孕方高剂量组PERK,BIP,CHOP蛋白表达显著降低(
P
<
0.01),CHOP mRNA表达水平显著升高(
P
<
0.01),而IRE-1和ATF6蛋白表达,PERK,IRE1,BIP,ATF6 mRNA表达及血清E
2
,P水平差异无统计学意义。
结论
2
补肾助孕方可以改善黄体功能不全,其机制可能与缓解内质网应激作用有关。
Objective
2
To investigate the regulatory mechanism of Bushen Zhuyun prescription(BSZYP) on endoplasmic reticulum stress (ERS) in rats with luteal phase defect (LPD) induced by mifepristone.
Method
2
Fifty SD rats were randomly divided into a blank group, a model group, a positive control group (dydrogesterone,0.02 g·kg
-1
), and low-(0.08 g·kg
-1
)and high-dose (0.24 g·kg
-1
) BSZYP groups. Western blot and Real-time fluorescence-based quantitative polymerase chain reaction (Real-time PCR) were used to detect the mRNA and protein expression levels of immunoglobulin binding protein (BIP), protein kinase R-like endoplasmic reticulum kinase (PERK), inositol-requiring enzyme 1 (IRE-1), activating transcription factor 6 (ATF6), and C/EBP homologous protein (CHOP). The enzyme-linked immunosorbent assay (ELISA) was used to detect the serum progesterone (P) and estradiol (E
2
) levels.
Result
2
Compared with the blank group, the model group showed the elevated protein expression of BIP, PERK, and CHOP (
P
<
0.01) and the dwindled mRNA expression of PERK and CHOP (
P
<
0.05), while no significant difference was observed in the protein expression of IRE-1 and ATF6, mRNA expression of IRE-1, BIP, and ATF6, and serum E
2
and P levels. Compared with the model group, the positive control group displayed diminished protein expression of CHOP (
P
<
0.01), while no significant difference was observed in the protein expression of PERK, IRE-1, BIP, and ATF6, mRNA expression of PERK, IRE-1, BIP, ATF6, and CHOP, and serum levels of E
2
and P. The protein expression of CHOP decreased (
P
<
0.01) and the mRNA expression of CHOP increased (
P
<
0.05) in the low-dose BSZYP group, while no significant difference was observed in the mRNA and protein expression of PERK, IRE-1, BIP, and ATF6, and serum E
2
and P levels. In the high-dose BSZYP group, the protein expression of PERK, BIP, and CHOP was down-regulated (
P
<
0.01), and the mRNA expression of CHOP was up-regulated (
P
<
0.01), while no significant difference was observed in the protein expression of IRE-1 and ATF6, mRNA expression of PERK, IRE-1, BIP, and ATF6, and serum E
2
and P levels.
Conclusion
2
BSZYP can treat LPD by relieving ERS.
冒韵东 , 刘嘉茵 . 不孕症诊治临床路径 [J]. 中国实用妇科与产科杂志 , 2013 , 29 ( 9 ): 690 - 693 .
邱雅琳 , 刘音吟 , 周惠芳 . 黄体功能不全性不孕症临证思路撷要 [J]. 北京中医药 , 2019 , 38 ( 7 ): 665 - 667 .
杨碧蓉 , 周华 . 寿胎丸加味联合黄体酮及低分子肝素治疗复发性流产的临床观察 [J]. 世界中医药 , 2019 , 14 ( 6 ): 1502 - 1506 .
刘冬娥 . 促排卵周期中的黄体支持 [J]. 中国实用妇科与产科杂志 , 2015 , 31 ( 1 ): 35 - 38 .
孙爱军 , 唐旭东 , 张巧利 , 等 . 卵巢储备功能降低不孕症中西医结合治疗的理论与临床试验研究探讨 [J]. 中国实验方剂学杂志 , 2019 , 25 ( 8 ): 148 - 157 .
李姣 , 林洁 . 龙胆泻肝汤治疗湿热瘀结型盆腔炎性不孕的疗效及作用机制 [J]. 中国实验方剂学杂志 , 2020 , 26 ( 6 ): 85 - 90 .
杨斌 , 周文清 , 陈莎 , 等 . 补肾助孕方对肾虚薄型子宫内膜大鼠内膜容受性的影响及机制研究 [J]. 世界中医药 , 2020 , 15 ( 17 ): 2565 - 2569 .
袁艺祯 , 周惠芳 , 蒋小飞 , 等 . 补肾助孕方对LPD模型大鼠卵巢SOD-2、GPX-1、P53表达水平的影响 [J]. 时珍国医国药 , 2019 , 30 ( 8 ): 1814 - 1816 .
LI L , ZHENG P , DEAN J . Maternal control of early mouse development [J]. Development , 2010 , 137 ( 6 ): 859 - 870 .
TAKAHASHI N , HARADA M , HIROTA Y , et al . A potential role for endoplasmic reticulum stress in progesterone deficiency in obese women [J]. Endocrinology , 2017 , 158 ( 1 ): 84 - 97 .
PARK H J , LEE D G , SEONG J B , et al . Peroxiredoxin I maintain luteal function by regulating unfolded protein response [J]. Reprod Biol Endocrinol , 2018 , 16 ( 1 ): 79 .
周伯如 , 戴佳轩 , 周惠芳 , 等 . 排卵后给予米非司酮对雌性大鼠动情周期、黄体功能的影响及意义 [J]. 山东医药 , 2017 , 57 ( 44 ): 32 - 35 .
刘贝 , 周惠芳 , 周伯如 , 等 . 补肾助孕方对大鼠垂体细胞促性腺激素的调节 [J]. 医学研究生学报 , 2018 , 31 ( 7 ): 703 - 708 .
王飞虹 , 谈勇 . 黄体功能不全性不孕中西医治疗进展 [J]. 吉林中医药 , 2015 , 35 ( 3 ): 319 - 324 .
ESENCAN E , KALLEN A , ZHANG M , et al . Translational activation of maternally derived mRNAs in oocytes and early embryos and the role of embryonic poly(A) binding protein (EPAB) [J]. Biol Reprod , 2019 , 100 ( 5 ): 1147 - 1157 .
PRASAD M , PAWLAK K J , BURAK W E , et al . Mitochondrial metabolic regulation by GRP78 [J]. Sci Adv , 2017 , doi: 10.1126/sciadv.1602038 http://dx.doi.org/10.1126/sciadv.1602038 .
XIONG Y , CHEN H , LIN P , et al . ATF6 knockdown decreases apoptosis,arrests the S phase of the cell cycle,and increases steroid hormone production in mouse granulosa cells [J]. Am J Physiol Cell Physiol , 2017 , 312 : C341 - C353 .
PARK S J , KIM T S , PARK C K , et al . hCG-induced endoplasmic reticulum stress triggers apoptosis and reduces steroidogenic enzyme expression through activating transcription factor 6 in Leydig cells of the testis [J]. J Mol Endocrinol , 2013 , 50 ( 2 ): 151 - 166 .
PARK H J , PARK S J , KOO D B , et al . Progesterone production is affected by unfolded protein response (UPR) signaling during the luteal phase in mice [J]. Life Sci , 2014 , 113 ( 1/2 ): 60 - 67 .
徐倩 , 许银丰 , 杨杰杰 , 等 . 内质网应激与细胞自噬的关系 [J]. 中国细胞生物学学报 , 2020 , 42 ( 8 ): 1489 - 1500 .
0
浏览量
23
下载量
2
CSCD
关联资源
相关文章
相关作者
相关机构
京公网安备11010802024621